Takehiro Miyasaka

Hollow-fiber blood-dialysis membranes: superoxide generation, permeation, and dismutation measured by chemiluminescence.

The interaction of blood with a material surface results in activation of the bodys humoral immune system and the generation of reactive oxygen species (ROS). It has recently become clear that ROS are central to the pathology of many diseases. In this study, we evaluated the superoxide generation, permeation, and dismutation in hollow-fiber dialysis membranes by using 2-methyl-6-p-methoxyphenylethynyl-imidazopyrazinone (MPEC) as a superoxide-reactive chemiluminescence producer and an optical fiber probe to detect the resulting chemiluminescence in the hollow fiber lumen. We measured the superoxide generated when bovine blood leukocytes were brought into contact with dialysis membranes. Superoxide permeation was determined by measuring MPEC chemiluminescence in the hollow fiber lumen using an optical fiber probe. Additionally, superoxide dismutation was evaluated by examining the difference in superoxide permeability for membranes with and without vitamin E coating. Superoxide generation varies for different membrane materials, depending on the membranes biocompatibility. Superoxide permeability depends on the diffusive permeability of membranes. No marked decrease in superoxide permeability was observed among membrane materials. The superoxide permeability of vitamin E-coated membrane was smaller than that of uncoated membrane. The antioxidant property of vitamin E-coated membranes is hence effective in causing superoxide dismutation.

Systemic nitric oxide production rate during hemodialysis and its relationship with nitric oxide-related factors.

Background/Aims:Nitric oxide (NO) plays a key role in the regulation of vascular tone and controls both local and systemic hemodynamics. Here, we estimated systemic NO production rates of hemodialysis (HD) patients, based on the time course of plasma concentration of nitrate (an oxidative end product of NO) and investigated possible roles of NO-related factors. Methods: We measured plasma concentrations of nitrate, L-arginine (a substrate of NO synthase: NOS), asymmetric dimethylarginine (ADMA, an endogenous NOS inhibitor), tetrahydrobiopterin (BH4, a NOS cofactor), dihydrobiopterin (BH2, an oxidized form of BH4) and oxidized low-density lipoprotein (oxyLDL; an index of oxidative stress) before and after 30-min and 4-hour HD (n = 10). Results:The time-averaged NO production rate during HD was estimated by fitting the time course of plasma nitrate concentration with a single-compartment model (4.00 ± 0.82 µmol/min, 4.99 ± 1.08 µmol/kg/h). The L-arginine/ADMA ratio (L-arginine availability) after 30-min HD showed a positive correlation with the NO production rate (p < 0.05). Conclusion: The systemic NO production rate during HD could be estimated by the single-compartment analysis. The L-arginine/ADMA ratio seems to play an important role in the regulation of the NO production during HD.

Direct Measurement of Nitric Oxide during Experimental Cardiopulmonary Bypass

We developed a system to measure nitric oxide (NO) concentration during cardiopulmonary bypass in anaesthetized pigs (n = 6). A T-shaped connector, attached to an NO sensor, was mounted in the extra-corporeal circuit at two measuring sites: proximal to the membrane oxygenator (venous side) and distal to the arterial line filter (arterial side). After performing a preliminary validation study, we measured plasma NO concentration before and during total cardiopulmonary bypass circulation (non-pulsatile flow 1.5 l/min) and without pulmonary ventilation. After establishing bypass, PaO2 was 318-393 mmHg; when PaO2 was decreased to 80-100 mmHg, plasma NO concentration in the arterial circuit fell by 39.2 ± 15.6 nM. There was no observable change in plasma NO concentration at the venous circuit. This new system could be useful in monitoring NO concentration during cardiac surgery with cardiopulmonary bypass, and for understanding the possible pathophysiological roles of hyper-nitric oxaemia in cardiopulmonary bypass-related cardiovascular complications.

Development of a contamination free 6 valve injector inline monitoring system for endotoxin measurement in dialysate.

Use of dialysate as supplement fluid in hemodiafiltration requires controlling contamination by endotoxin of the dialysate. We thus aimed at developing an endotoxin monitoring system with complete exclusion of endotoxin contamination for simple, easy, and accurate measurement of endotoxin concentration in dialysate. In the present study, we used a 6 valve injector along with a high performance liquid chromatogram system. This new system showed a sensitivity of approximately 1 endotoxin units (EU)/L in the range of 0 to 30 EU/L endotoxin in dialysate and no trace of endotoxin contamination. In conclusion, the new endotoxin monitoring system showed high sensitivity and reproducibility, with easy operation.